Levengood Alexis L, Hohwieler Katrin, Powell Daniel, Cristescu Romane H
Detection Dogs for Conservation, School of Science, Technology, and Engineering University of the Sunshine Coast Sippy Downs Queensland Australia.
Marine and Terrestrial Megafauna Research Cluster University of the Sunshine Coast Sippy Downs Queensland Australia.
Ecol Evol. 2025 Jul 9;15(7):e71755. doi: 10.1002/ece3.71755. eCollection 2025 Jul.
Recent advances in DNA sequencing and genotyping technologies are rapidly building our capacity to address ecological, evolutionary, and conservation questions for wildlife. However, wildlife genetic research increasingly relies on samples containing low quantities and quality of DNA, such as non-invasive, archival, and environmental DNA. These samples present unique methodological challenges; for samples collected in the wild, it is important to understand the impact of environmental exposure time or sample 'age' on DNA quality and downstream genetic analyses. Here, we aged koala () scats under natural conditions and quantified DNA degradation. We assessed the effect of age on genetic data quality by measuring the proportion of missing single nucleotide polymorphism (SNP) data using DArTCap, a targeted genotyping methodology hypothesised to tolerate degraded DNA. Contrary to other studies, we found koala scat age was not a significant predictor of genotyping quality (i.e., rate of missing SNP calls) in the first 10 days of environmental exposure. We yielded high quality data from 10-day-old scats, but also low-quality data from fresh scats. This study is the first to investigate the effect of scat age on genotyping success using a targeted approach, and DArTCap specifically. These findings support the use of targeted genotyping (such as DArTCap) from scats and provide insights for future research using DNA from non-invasive samples. Targeted genotyping may extend the timeframe, from which accurate data can be obtained from non-invasive samples, increasing potential sample sizes and enhancing our ability to address important questions in population ecology, conservation genetics, and population management.
DNA测序和基因分型技术的最新进展正在迅速提升我们解决野生动物生态、进化和保护问题的能力。然而,野生动物遗传学研究越来越依赖于DNA含量和质量较低的样本,例如非侵入性样本、存档样本和环境DNA样本。这些样本带来了独特的方法学挑战;对于在野外采集的样本,了解环境暴露时间或样本“年龄”对DNA质量和下游基因分析的影响非常重要。在这里,我们在自然条件下对考拉粪便进行了年代测定,并对DNA降解进行了量化。我们使用DArTCap(一种假定能够耐受降解DNA的靶向基因分型方法)测量缺失单核苷酸多态性(SNP)数据的比例,以此评估样本年龄对遗传数据质量的影响。与其他研究相反,我们发现考拉粪便年龄在环境暴露的前10天并不是基因分型质量(即SNP调用缺失率)的显著预测指标。我们从10日龄的粪便中获得了高质量的数据,但也从新鲜粪便中获得了低质量的数据。本研究首次使用靶向方法,特别是DArTCap,来研究粪便年龄对基因分型成功率的影响。这些发现支持了从粪便中使用靶向基因分型(如DArTCap),并为未来使用非侵入性样本中的DNA进行研究提供了见解。靶向基因分型可能会延长从非侵入性样本中获取准确数据的时间框架,增加潜在样本量,并增强我们解决种群生态学、保护遗传学和种群管理中重要问题的能力。